Effect of water-soluble reduced chitosan on Streptococcus mutans, plaque regrowth and biofilm vitality

  • K. Bae
  • E. J. Jun
  • S. M. Lee
  • D. I. Paik
  • J. B. Kim
Original Article


The purpose of the present study was to examine the effects of a newly developed water-soluble reduced chitosan on Streptococcus mutans, plaque regrowth, and biofilm vitality. A 1.0%, water-soluble reduced chitosan, with pH ranging from 6.0 to 6.5, molecular weights between 3,000 and 5,000 Da, and 70% degree of deacetylation, was used. To determine antibacterial and antiplaque potency of chitosan, minimal inhibitory concentrations (MICs) for S. mutans and S. sanguinis (formerly S. sanguis), short-term exposure to S. mutans, and clinical trial of plaque regrowth and biofilm vitality were conducted. Twelve dental students volunteered to participate in the 6-week, double blind, randomized clinical trial using the classical 4-day plaque regrowth design. The MIC of water-soluble reduced chitosan for S. mutans was 1.25 g/l. While the cells exposed to distilled water (DW) grew rapidly, with a maximum turbidity reached by 16 h postinoculation, S. mutans exposed to chitosan (5.0 g/l) exhibited a substantial delay in growth and reached a maximum turbidity by 32 h postinoculation. The chitosan solution reduced the plaque index and the vitality of the plaque flora significantly when compared to DW, but this was less than the reductions found with the positive control of 0.1% chlorhexidine solution. The water-soluble reduced chitosan exhibited potent antibacterial effect on S. mutans, and displayed a significant antibacterial and plaque-reducing action during the 4-day plaque regrowth.


Water-soluble reduced chitosan Streptococcus mutans Plaque regrowth Biofilm vitality Minimal inhibitory concentrations 



The statistical advice of Choongrak Kim (Pusan National University, Busan, South Korea) is gratefully acknowledged.


  1. 1.
    Addy M, Moran J, Griffiths AA, Wills-Wood NJ (1985) Extrinsic tooth discoloration by metals and chlorhexidine. I. Surface protein denaturation or dietary precipitation? Br Dent J 159:281–285CrossRefPubMedGoogle Scholar
  2. 2.
    al-Delaimy KS, Ali SH (1970) Antibacterial action of vegetable extracts on the growth of pathogenic bacteria. J Sci Food Agric 21:110–112PubMedCrossRefGoogle Scholar
  3. 3.
    Arweiler NB, Henning G, Reich E, Netuschil L (2002) Effect of an amine–fluoride–triclosan mouthrinse on plaque regrowth and biofilm vitality. J Clin Periodontol 29:358–363CrossRefPubMedGoogle Scholar
  4. 4.
    Baker PJ, Coburn RA, Genco RJ, Evans RT (1987) Structural determinants of activity of chlorhexidine and alkyl bisbiguanides against the human oral flora. J Dent Res 66:1099–1106PubMedGoogle Scholar
  5. 5.
    Binney A, Addy M, McKeown S, Everatt L (1996) The choice of controls in toothpaste studies. The effect of a number of commercially available toothpastes compared to water on 4-day plaque regrowth. J Clin Periodontol 23:456–459CrossRefPubMedGoogle Scholar
  6. 6.
    Brecx M, Netuschil L, Reichert B, Schreil G (1990) Efficacy of Listerine, Meridol and chlorhexidine mouthrinses on plaque, gingivitis and plaque bacteria vitality. J Clin Periodontol 17:292–297CrossRefPubMedGoogle Scholar
  7. 7.
    Brecx M (1997) Strategies and agents in supragingival chemical plaque control. Periodontol 2000 15:100–108PubMedCrossRefGoogle Scholar
  8. 8.
    Choi BK, Kim KY, Yoo YJ, Oh SJ, Choi JH, Kim CY (2001) In vitro antimicrobial activity of a chitooligosaccharide mixture against Actinobacillus actinomycetemcomitans and Streptococcus mutans. Int J Antimicrob Agents 18:553–557CrossRefPubMedGoogle Scholar
  9. 9.
    Choi HJ, Lee HS, Her S, Oh DH, Yoon SS (1999) Partial characterization and cloning of leuconocin J, a bacteriocin produced by Leuconostoc sp. J2 isolated from the Korean fermented vegetable Kimchi. J Appl Microbiol 86:175–181CrossRefPubMedGoogle Scholar
  10. 10.
    Decker EM, von Ohle C, Weiger R, Wiech I, Brecx M (2005) A synergistic chlorhexidine/chitosan combination for improved antiplaque strategies. J Periodontal Res 40:373–377CrossRefPubMedGoogle Scholar
  11. 11.
    Drake DR, Wefel JS, Dunkerson D, Hogle K (1993) The antimicrobial activity of Prevention mouthrinse. Am J Dent 6:239–242PubMedGoogle Scholar
  12. 12.
    Flotra L, Gjermo P, Rolla G, Waerhaug J (1971) Side effects of chlorhexidine mouth washes. Scand J Dent Res 79:119–125PubMedGoogle Scholar
  13. 13.
    Fujiwara M, Hayashi Y, Ohara N (2004) Inhibitory effect of water-soluble chitosan on growth of Streptococcus mutans. New Microbiol 27:83–86PubMedGoogle Scholar
  14. 14.
    Holme KR, Perlin AS (1997) Chitosan N-sulfate. A water-soluble polyelectrolyte. Carbohydr Res 302:7–12CrossRefPubMedGoogle Scholar
  15. 15.
    Ikinci G, Senel S, Akincibay H, Kas S, Ercis S, Wilson CG, Hincal AA (2002) Effect of chitosan on a periodontal pathogen Porphyromonas gingivalis. Int J Pharm 235:121–127CrossRefPubMedGoogle Scholar
  16. 16.
    Järvinen H, Tenovuo J, Huovinen P (1993) In vitro susceptibility of Streptococcus mutans to chlorhexidine and six other antimicrobial agents. Antimicrob Agents Chemother 37:1158–1159PubMedGoogle Scholar
  17. 17.
    Johnson MG, Vaughn RH (1969) Death of Salmonella typhimurium and Escherichia coli in the presence of freshly reconstituted dehydrated garlic and onion. Appl Microbiol 17:903–905PubMedGoogle Scholar
  18. 18.
    Keene HJ, Shklair IL (1974) Relationship of Streptococcus mutans carrier status to the development of carious lesions in initially caries free recruits. J Dent Res 53:1295PubMedGoogle Scholar
  19. 19.
    Kim JH (1995) Inhibition of Listeria monocytogenes by bacteriocin from lactic acid bacteria isolated from kimchi. Agric Chem Biotechnol 38:302–307Google Scholar
  20. 20.
    Kornman KS (1985) Antimicrobial agents. National Institute of Dental Research, Bethesda, pp 150–194Google Scholar
  21. 21.
    Löe H and Silness J (1963) Periodontal disease in pregnancy: I. Prevalence and severity. Acta Odontol Scand 21:533–551PubMedCrossRefGoogle Scholar
  22. 22.
    Miyazaki S, Nakayama A, Oda M, Takada M, Attwood D (1994) Chitosan and sodium alginate based bioadhesive tablets for intraoral drug delivery. Biol Pharm Bull 17:745–747PubMedGoogle Scholar
  23. 23.
    Moran J, Addy M, Newcombe RG, Marlow I (2000) A study to assess the plaque inhibitory activity of a new triclosan mouthrinse formulation. J Clin Periodontol 27:806–809CrossRefPubMedGoogle Scholar
  24. 24.
    Netuschil L, Reich E, Brecx M (1989) Direct measurement of the bactericidal effect of chlorhexidine on human dental plaque. J Clin Periodontol 16:484–488CrossRefPubMedGoogle Scholar
  25. 25.
    Netuschil L, Weiger R, Preisler R, Brecx M (1995) Plaque bacteria counts and vitality during chlorhexidine, meridol and listerine mouthrinses. Eur J Oral Sci 103:355–361PubMedCrossRefGoogle Scholar
  26. 26.
    Newcombe RG (1992) Crossover trials comparing several treatments. J Clin Periodontol 19:785–787CrossRefPubMedGoogle Scholar
  27. 27.
    Newcombe RG, Addy M, McKeown S (1995) Residual effect of chlorhexidine gluconate in 4-day plaque regrowth crossover trials, and its implications for study design. J Periodontal Res 30:319–324PubMedCrossRefGoogle Scholar
  28. 28.
    Perneger TV (1998) What's wrong with Bonferroni adjustments. BMJ 316:1236–1238PubMedGoogle Scholar
  29. 29.
    Reichart P, Gehring F (1984) Streptococcus mutans and caries prevalence in Lisu and Karen of northern Thailand. J Dent Res 63:56–58PubMedGoogle Scholar
  30. 30.
    Rölla G, Kjærheim V, Waaler SM (1994) The role of antiseptics in primary prevention. In: Lang NP, Karring T (eds) Proceedings of the second European Workshop in Periodontology. Quintessence, LondonGoogle Scholar
  31. 31.
    Schiött CR, Loe H, Jensen SB, Kilian M, Davies RM, Glavind K (1970) The effect of chlorhexidine mouthrinses on the human oral flora. J Periodontal Res 5:84–89PubMedCrossRefGoogle Scholar
  32. 32.
    Seo H, Shoji A, Itoh Y, Kawamura M, Sakagami Y (1994) Antibacterial fiber blended with chitosan. In: Karnicki ZS, Brzeski MM, Bykowski PJ, Wojtasz-Pajak A (eds) Chitin world. Wirtschaftverlag NV-Verlag für neue Wissenschaft GmbH, Germany, pp 623–631Google Scholar
  33. 33.
    Silness J, Löe H (1964) Periodontal disease in pregnancy (II). Correlation between oral hygiene and periodontal condition. Acta Odontol Scand 22:121–135PubMedCrossRefGoogle Scholar
  34. 34.
    Sugimoto M, Morimoto M, Sashiwa H, Saimoto H, Shigemasa Y (1998) Preparation and characterization of water-soluble chitin and chitosan derivatives. Carbohydr Polym 36:49–59CrossRefGoogle Scholar
  35. 36.
    Tarsi R, Muzzarelli RA, Guzman CA, Pruzzo C (1997) Inhibition of Streptococcus mutans adsorption to hydroxyapatite by low-molecular-weight chitosans. J Dent Res 76:665–672PubMedCrossRefGoogle Scholar
  36. 35.
    Tarsi R, Corbin B, Pruzzo C, Muzzarelli RA (1998) Effect of low-molecular-weight chitosans on the adhesive properties of oral streptococci. Oral Microbiol Immunol 13:217–224PubMedCrossRefGoogle Scholar
  37. 37.
    Tsai GJ, Zhang SL, Shieh PL (2004) Antimicrobial activity of a low-molecular-weight chitosan obtained from cellulase digestion of chitosan. J Food Prot 67:396–398PubMedGoogle Scholar
  38. 38.
    Turesky S, Gilmore ND, Glickman I (1970) Reduced plaque formation by the chloromethyl analogue of victamin C. J Periodontol 41:41–43PubMedGoogle Scholar
  39. 39.
    Yang TC, Chou CC, Li CF (2005) Antibacterial activity of N-alkylated disaccharide chitosan derivatives. Int J Food Microbiol 97:237–245CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag 2006

Authors and Affiliations

  • K. Bae
    • 1
  • E. J. Jun
    • 1
  • S. M. Lee
    • 1
  • D. I. Paik
    • 2
  • J. B. Kim
    • 1
  1. 1.Department of Preventive and Community Dentistry, College of DentistryPusan National UniversitySo-guSouth Korea
  2. 2.Department of Preventive and Public Health Dentistry, College of DentistrySeoul National UniversityChongro-guSouth Korea

Personalised recommendations